Reagent mixer and fluid control devices

ABSTRACT

An apparatus for preparing a reagent solution includes an enclosure and a container disposed within the enclosure. The container defines an internal cavity having a compressible volume and defines a passage providing fluidic communication between the internal cavity and the exterior of the container. Optionally, a compressible member is disposed within the internal cavity. A reagent is disposed within the internal cavity.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application is a divisional application under 35 U.S.C. § 120 ofpending U.S. application Ser. No. 14/739,974 filed Jun. 15, 2015, whichapplication claims benefit under 35 U.S.C. § 119(e) of U.S. ProvisionalApplication No. 62/012,685 filed Jun. 16, 2014. The entire contents ofthe aforementioned applications are incorporated herein.

TECHNICAL FIELD

This application relates to the field of mixing fluids, and inparticular, an apparatus and method for preparing a reagent solution.

BACKGROUND

In certain devices, reagents are delivered to produce a reaction productmonitored by sensors. However, there are currently no adequate solutionsthat allow for efficient storage and transportation of reagents thatalso limit user handling and interaction, including potentialcontamination, in preparation of the mixed reagents. Thorough andconsistent mixing of a reagent in solution is also desirable. In view ofthe above, it would be advantageous to have a device for preparing areagent solution which overcame the deficiencies of current approaches.

SUMMARY

In an example, a reagent is disposed in a container including acompressible volume and one or more passages. The pressure of fluidexternal to the container is increased to drive fluid through thepassage and into the container, compressing the compressible volume.When the pressure of the fluid is decreased, the compressible volumedecompresses, driving fluid and the reagent out of the container. Thecontainer can be disposed in an enclosure, such as a flexible enclosure.One or more of the enclosures including the container can be disposed ina cartridge, which can be pressurized.

BRIEF DESCRIPTION OF THE DRAWINGS

The present disclosure may be better understood, and its numerousfeatures and advantages made apparent to those skilled in the art byreferencing the accompanying drawings.

FIG. 1 is a perspective view describing an exemplary reagent storageapparatus.

FIG. 2 is a perspective view describing an example container.

FIG. 3 is a cross-sectional perspective view describing an examplecontainer.

FIG. 4 is an exploded schematic view describing an example container.

FIG. 5 is a detailed perspective view describing an example container.

FIG. 6 is a perspective view describing an exemplary reagent storageapparatus.

FIG. 7 is a perspective view describing an example container.

FIG. 8 is a cross-sectional perspective view describing an examplecontainer.

FIG. 9, FIG. 10, FIG. 11, FIG. 12, and FIG. 13 include illustrations ofan exemplary cartridge for enclosing one or more enclosures.

FIG. 14 is a block diagram describing an example system.

The use of the same reference symbols in different drawings indicatessimilar or identical items.

DETAILED DESCRIPTION

Reference will now be made in detail to implementations, examples ofwhich are illustrated in the accompanying drawings. In the followingdetailed description, numerous specific details are set forth in orderto provide a sufficient understanding of the subject matter presentedherein. But it will be apparent to one of ordinary skill in the art thatthe subject matter may be practiced without these specific details.Moreover, the particular implementations described herein are providedby way of example and should not be used to limit the scope of theinvention to these particular implementations. In other instances,well-known structures and components have not been described in detailso as not to unnecessarily obscure aspects of the implementations of theinvention.

FIG. 1 is a perspective view describing an exemplary reagent storageapparatus 100. In an example, the reagent storage apparatus 100 caninclude an enclosure 110. A container 120 is disposed within theenclosure 110. In an example, the enclosure 110 can be a flexibleenclosure. A flexible enclosure, such as a sealable flexible bagenclosure, can be pressurized and depressurized by externally applyingpressure, such by applying pressurized gas to the external surfaces ofthe flexible enclosure. Alternatively, the enclosure can be rigid sothat externally applied gas pressure does not readily translate topressure of fluid within the enclosure 110.

The reagent storage apparatus 100 can also include a fluid port 130coupled to a fitting 160 attached to the enclosure 110 to provide fluidaccess to the interior of the enclosure 110. The fluid port 130 can becoupled to the fitting 160 to seal the enclosure 110 from an exteriorenvironment after insertion of the container 120. The enclosure 110 canbe, for example, thermosealed to itself and the fitting 160, exceptwhere otherwise sealed by the fluid port 130.

The container 120 can include one or more arms 140 and a flange 150. Thearms 140 can position the container 120 within the enclosure 110, suchas approximately centrally, to disperse a reagent within the enclosure100 evenly. The flange 150 can be provided for convenient assembly ofthe container 120. In an example, the arm 140 is flexible. For example,the arm 140 can be formed of wire or a polymeric material.Alternatively, the arm 140 can be rigid. Alternatively, the arm 140 andthe flange 150 are not limited to those illustrated in FIG. 1 and caninclude a structure that positions the container within a predeterminedlocation or orientation within the enclosure 110. The container 120 canbe directly or indirectly connected to the fluid port 130 oralternatively positioned a suitable distance away from the fluid port130 as show in FIG. 1. The sealed enclosure 100 including the enclosure110 and container 120 provides simplified storage and transportation ofa reagent within the container 120.

FIG. 2 is a perspective view describing an example container 200. Thecontainer 200 can include a first portion 210 and a second portion 220coupled to the first portion 210. Elements such as an optionalcompressible member and a reagent can be inserted within the container200 prior to connecting the first portion 210 to the second portion 220.In an example, the second portion 220 can be a cap that slides over orotherwise covers part of the first portion 210 to form the internalcavity. In another example, the second portion 220 can be an insert thatslides into the first portion 210. The second portion 220 can beconnected to the first portion 210 by any suitable attachment mechanismincluding screwing the second portion 220 onto the first portion 210 orvice versa, a locking mechanism, adhesive, or any other suitableattachment mechanism.

In an example, the internal cavity defines a compressible volume. Thecompressible volume compresses in response to fluid pressure and doesnot dissipate or leave the internal cavity of the container 200. Thecompressible volume can include a compressible gas volume or can be acompressible member, such as a resilient polymer or foam.

The container 200 can define a passage 230 providing fluidiccommunication between an internal cavity of the container 200 and anexterior of the container 200. In an example, one or more passages 230can be defined through to the internal cavity. Such passages 230 can bedrilled through the first or second portions of the container 200. Inanother example, the second portion 220 can include the passage 230 orcan include a slot extending beyond a region at which the second portion220 engages the first portion 210, thus forming the passage 230.

One or more arms 240 can be coupled to the first portion 210 to positionthe container 200 as desired within an enclosure. A flange 250 can becoupled to the second portion 220 to assist with applying the secondportion 220 to the first portion 210 or to position the container 200away from a bottom of the enclosure.

FIG. 3 is a cross-sectional perspective view describing an examplecontainer 300. The container 300 defines an internal cavity 320 anddefines a passage 330 providing fluidic communication between theinternal cavity 320 and the exterior of the container 300. The passage330 can be drilled through the container 300. In another example, a capor insert can include a slot that extends beyond a region engaging thecontainer 300 and that forms the passage 330. The container 300 caninclude a first portion 310 and a second portion 350 coupled to thefirst portion 310 that allows elements such as a compressible member 340and reagent to be inserted into the internal cavity 320 of the container300.

The internal cavity 320 defines a compressible volume. The compressiblevolume is a volume that compresses in response to pressure to match thepressure, and can expand in response to depressuring, providing acounter force on fluid pressure. In an example, the compressible volumeincludes a compressible gas that compresses to match the pressure offluid entering the internal volume without dissipating or exiting theinternal cavity and in response to a depressurization of the fluidpushes the fluid out of the internal cavity 320. Optionally, thecompressible volume can include a compressible member 340. Thecompressible member 340 is compressible under pressurization and upondepressurization, substantially returns to its previous form. Forexample, the compressible member 340 can be a foam material. Inparticular, the compressible member 340 can be a closed-cell foam ofelastomeric material. In an example, the compressible member can includepolyurethane foam.

In an example, the reagent can be disposed within the second portion350. The reagent can be a lyophilized nucleotide or an analog thereof.In another example, the reagent is a solution absorbed on a porousmetal, ceramic, or polymeric sponge-like material or frit. Optionally,the reagent solution can be frozen. In an alternative example, thereagent can include a pH-adjusting reagent, such as an acid or base.

One or more arms 360 can be coupled to the first portion 310 to positionthe container 300 as desired within an enclosure. A flange 370 or othersuitable appendage can be coupled to the second portion 350 to assistwith engaging the second portion 350 with the first portion 310 or toposition the container 300 within the enclosure.

FIG. 4 is an exploded schematic view describing an example container400. The container 400 can include a first portion 410 and a secondportion 420 (e.g., an insert) coupled to the first portion 410, allowingelements such as an optional compressible member 430 to be insertedwithin the container 400. The second portion 420 can be secured to thefirst portion 410 by sliding or screwing into the first portion 410. Oneor more flexible arms 440 can be coupled to the first portion 410 toposition the container 400 within an enclosure. A flange 450 can becoupled to the second portion 420 to assist with engaging the secondportion 420 and the first portion 410 or to position the container 400within the enclosure.

In an example, the second portion 420 can define one or more slots 460.The first portion 410 and the second portion 420 can engage so as toleave a portion of the one or more slots 460 exposed, providing one ormore passages between the internal cavity of the container 400 and theexterior of the container 400.

FIG. 5 is a detailed perspective view describing an example container500. The detailed view of the container 500 can define a passage 510providing fluidic communication between an internal cavity of thecontainer 500 and an exterior of the container 500. An end of thecontainer 500 includes a fitting 530 to receive an insert 540. Theinsert 540 includes a hole or slot not covered when the insert 540 isapplied to the fitting, forming the passage 510. Alternatively, theinsert 540 can include a hole, notch, mesh, pores or any other suitablefeature for providing fluid communication to the fitting 530. A flange520 can be coupled to the container 500 to allow control of the insert540 as it is applied to the fitting 530 and to position the container200 (as illustrated in FIG. 2) within an enclosure.

FIG. 6 is a perspective view describing an exemplary reagent storageapparatus 600. The reagent storage apparatus 600 includes an enclosure610. A container 620 is disposed within the enclosure 610. The enclosure610 can be a flexible enclosure as described above. For example, theflexible enclosure can be a sealable flexible bag enclosure that can bepressurized and depressurized externally via fluid pressure or gaspressure. Alternatively, the enclosure 610 can be a rigid enclosure. Theenclosure 610 can sealably engage a seal structure 670, such as afitting, having a bore 680, such as a central bore. The container 620can be coupled to an arm 640, which can be coupled to a fluid port 630,and inserted through the bore 680 of the fitting 670.

The fluid port 630 provides fluid access to the interior of theenclosure 610 through the bore 680. The fluid port 630 can be coupled tothe seal structure or fitting 670 of the reagent storage apparatus 600from an exterior environment after inserting the container 620. The arm640 couples the container 620 to the fluid port 630 to position thecontainer 620, for example, approximately centrally within the enclosure610 to disperse a reagent within the enclosure 610 evenly. The arm 640,the container 620, and the fluid port 630 can be a single integratedpiece. In an example fluid flows through the fluid port 630 and throughthe bore 680 of the fitting 670 into the enclosure 610, optionally alongthe arm 640. The arm 640 can position the container 620 with or withouta flange 650.

FIG. 7 is a perspective view describing an example container 700. Thecontainer 700 can include a first portion 710 and a second portion 720coupled to the first portion 710 allowing elements such as an optionalcompressible member to be inserted within the container 700. A fluidport 730 is coupled to the container 700 and provides fluid access to anenclosure into which the container 700 is inserted. An arm 740 cancoupled the first portion 710 and the fluid port 730 to position thecontainer 700 within an enclosure.

In an example, the second portion 720 is an insert to engage the firstportion 710. In another example, the second portion 720 forms a cap tocover an end of the first portion 710. Fluid can flow through theopening 770 of the port 730 and to an opening 760 along the arm 740. Thefluid port can include a gasket to facilitate sealing. A flange 750 canbe coupled to the second portion 720.

FIG. 8 is a cross-sectional perspective view describing an examplecontainer 800. The container 800 defines an internal cavity 820 anddefines a passage 830 providing fluidic communication between theinternal cavity 820 and the exterior of the container 800. The container800 can include a first portion 810 and a second portion 850 coupled tothe first portion 810 defining a compressible volume. In an example,elements, such as a compressible member 840 and reagent, can be insertedinto the internal cavity 820 of the container 800.

In an example, the second portion 850 is a cap to apply over an end ofthe first portion 810. In another example, the second portion 850 is aninsert to apply to a fitting of the first portion 810. In an example, apassage 830 is formed in the second portion 850, for example, as a holeor a slot.

The reagent can be disposed within the second portion 850. The reagentcan be a lyophilized nucleotide or an analog thereof. In anotherexample, the reagent can be a nucleotide solution absorbed by a porousmetallic, ceramic or polymeric sponge or frit. In a further example, thereagent can be frozen. In an additional example, the reagent can includea pH-adjusting reagent, such as an acid or a base.

A fluid port 860 is coupled to the container 800 and provides fluidaccess to an enclosure in which the container 800 is inserted. Forexample, fluid entering opening 890 can pass through passages 895 andinto an enclosure. An arm 870 can be coupled to the first portion 810and the fluid port 860. A flange 880 can be coupled to the secondportion 850 to also position the container 800 within the enclosure.

The reagent storage apparatus can be inserted into a case or cartridgehaving a cavity. In an example, pressure can be varied within the cavityto change the pressure of liquid within the flexible enclosure and thus,influence the pressure within the container. Alternatively, pressure canbe applied through the opening 890 and internal to the enclosure. In anexample, one or more of the enclosures can be incorporated into thecase. The case can define one or more pressure chambers in whichpressure can be applied and relieved from the enclosures.

In a particular example illustrated in FIG. 9, a cartridge or case 900includes a lid 902 and a body 904. The lid 902 can receive the fluidports (906, 908, 910, 912, or 914) of containers inserted into flexibleenclosures. The container can include different reagents. For example,each container can include a nucleotide or can include a pH-adjustingreagent. The lid 902 can also include a port 916 for providingpressurized gas or relieving the pressure, controlling the pressureoutside of each of the enclosures and thereby controlling pressurewithin the enclosure. The walls of the base 904 and the lid 902 can beconfigured to permit pressurizing a cavity within the cartridge 900, forexample, with pressurized gas or air. In an example, the cartridge 900can be labeled with a bar code or radio frequency identification (RFID)tag.

As illustrated in FIG. 9 and FIG. 10, the lid 902 can include accessports 918 and 920 for applying gas or air through a scrubber cartridge.In particular, system can utilize external air, applying the externalair through the port 918 and receiving a cleaned gas or air through theport 920. In particular, the scrubber cartridge can include absorbentmaterials for capturing carbon dioxide or water. Carbon dioxide can beremoved from air to prevent acidification of liquid components whencarbon dioxide diffuses into the enclosures or when the air is used inother parts of the system.

In a further example, the lid 902 can also include alignment features924 or 926. Such alignment features can be used to align access to theports (906, 908, 910, 912, 914, 916, 918, or 920) with a manifold tolimit damage to the manifold or provide for adequate engagement betweenthe manifold and the case 900.

As illustrated in FIG. 11, FIG. 12, and FIG. 13, the body 904 can defineindividual cavities 1126 into which each enclosure 1128 is placed andthe nucleotide container 1130 is inserted. In an example, each enclosure1128 is disposed within individual cavities 1126 and each container 1130is applied through the lid 902, engaging the lid 902 at the fluid portof the container 1130. A fitting 1134 of the enclosures 1128 can engagethe lid 902.

The lid 902 can define a headspace that provides communication betweenthe pressurized gas input port 916 and each of the cavities 1126.Alternatively, the cavity can be an open cavity absent individualizedcavities 1126 and provide a single cavity to which pressurized gas canbe applied to apply pressure to the enclosures 1128. As illustrated inFIG. 12, the body 904 can include a chamber 1232 to receive a scrubbercartridge, for example, for removing carbon dioxide from the air.

In a top view, as illustrated in FIG. 13, the body 904 includesindividualized cavities 1126. In addition, the body can include a sealstructure 1340 to isolate the scrubber cartridge input and output fromthe pressure of the rest of the body 904. In addition, an internal seal1342 can be utilized to isolate the input pressure of air entering thescrubber cartridge from the output pressure of air leaving scrubbercartridge. Further, the body 904 can include a seal structure 1344 toengage an opposing seal structure on the lid 902 to provide an isolatedinterior space including the cavities that can be pressurized ordepressurized.

The containers can include nucleotide reagents or other reagents. Inparticular, individual containers within the cartridge system caninclude one of four nucleotides. The system can also include a containerwithin an enclosure that includes pH-adjusting reagents. In a particularexample, the cartridge includes containers and enclosures incorporatingeach of the four nucleotides (A, G, C, or T) and optionally, apH-adjusting reagent container. In an example, the reagents are in driedform. For example, lyophilized nucleotides can be stored within thecontainer. In another example, a reagent solution can be absorbed withina porous metallic, ceramic, or polymeric sponge-like material or frit.In a further example, the reagent solution can be frozen either within acontainer or within the porous sponge-like material into which thereagent solution is absorbed.

The enclosures described herein can be applied to prepare a reagentsolution. Assembly of the enclosure includes inserting a container intothe enclosure and sealing the container within the enclosure with afluid port. One or more enclosures can be further secured into a volumeof a case, where the case includes a gas port for providing external gaspressure to the secured enclosures. The enclosures can be inserted intoa case as a final assembly step or at a point just prior to mixing thatprovides flexibility of in the selection of reagents.

Alternatively, the enclosure can be secured to the lid prior toinserting the containers including reagent. The reagent containers canbe inserted through the lid and the fluid port of the containers canengage the lid. The lid can be secured to the base following securingthe enclosures to the lid or following inserting the containers throughthe lid into the enclosures.

The pressurization and depressurization of the fluid within theenclosures are controlled by increasing and decreasing the gas pressureof the volume of the case via the gas port.

A method for preparing a reagent solution includes filling an enclosure,such as any of the enclosures described herein including a container anda reagent, with a predetermined amount of fluid through a fluid port ofthe enclosure. The fluid within the enclosure is then pressurized suchthat fluid flows into the internal cavity of the container through apassage of the container. The fluid can be pressurized directly througha port. In another example, the fluid can be pressurized by applyingexternal pressure to the enclosure, for example, using gas or otherfluidic pressure. The pressurization compresses the compressible volumeor member within the internal cavity of the container while the fluidfills a portion of the volume of the internal cavity.

For example, the fluid flows into the internal cavity of the containerand compresses the compressible volume or member until the pressurewithin the internal cavity and exerted on the compressible volume ormember is approximately equal to a pressure within the enclosure andexternal to the container.

After reaching a predetermined pressure, the fluid within the enclosureis depressurized. The compressible volume or member decompresses so asto expand and eject the fluid and reagent from the internal cavity intothe enclosure outside of the container. The mixture of reagent and fluidejected from the passage creates eddy currents and turbulence within thebag enclosure sufficient to mix the reagent with fluid. Upondepressurization, a pressure within the internal cavity as imposed bythe compressible volume or member is greater than a pressure within theenclosure and external to the container. The fluid and the reagent ejectfrom the internal cavity through the passage until the pressure withinthe internal cavity is approximately equal to a pressure within theenclosure and external to the container to provide a well-mixed reagentsolution.

Pressurization can be performed by increasing a gas pressure external toa flexible enclosure. In one implementation, the enclosure can bedisposed within a case. Pressure within the enclosure can be controlledby increasing/decreasing a gas pressure within the case and external tothe enclosure. Proper mixing of the reagent and fluid can beaccomplished through repeated cycles of pressurization anddepressurization. After mixing is completed, the fluid and the reagentare released through the fluid port of the enclosure.

Exemplary enclosures are useful as reagent reservoirs in biologicalprocesses where multiple reagents are delivered to one or more reactorsor reaction sites. The reaction sites can be monitored by chemical,electrical or optical sensors. Exemplary systems include methods andapparatuses for carrying out DNA sequencing, and in particular, pH-basedDNA sequencing. For example, in pH-based DNA sequencing, baseincorporations are determined by measuring hydrogen ions that aregenerated as natural byproducts of polymerase-catalyzed extensionreactions. DNA templates each having a primer and polymerase operablybound are loaded into reaction chambers or microwells, after whichrepeated cycles of deoxynucleoside triphosphate (dNTP) addition andwashing are carried out. Such templates are typically attached as clonalpopulations to a solid support, such as a microparticle, bead, or thelike, and such clonal populations are loaded into reaction chambers. Ineach addition step of the cycle, the polymerase extends the primer byincorporating added dNTP when the next base in the template is thecomplement of the added dNTP. If there is one complementary base, thereis one incorporation, if two, there are two incorporations, if three,there are three incorporations, and so on. With each such incorporationthere is a hydrogen ion released, and collectively a population oftemplates releasing hydrogen ions causing very slight changes to thelocal pH of the reaction chamber which is detected by an electronicsensor. In addition to sequencing, the device herein can be useful forother biological instruments that require fluid storage or delivery.

FIG. 14 diagrammatically illustrates a system employing an enclosure1414 that is a reagent reservoir, for example, for carrying out pH-basednucleic acid sequencing. Each electronic sensor of the apparatusgenerates an output signal. The fluid circuit permits multiple reagentsto be delivered to the reaction chambers.

In FIG. 14, the system 1400 includes a fluidics circuit 1402 connectedto at least two reagent reservoirs 1414, to a waste reservoir 1420, andto a biosensor 1434 by fluid pathway 1432 that connects fluidics node1430 to inlet 1438 of biosensor 1434. The prepared and mixed reagentsolution from reservoirs 1414 can be driven to fluidic circuit 1402 by avariety of methods including pressure, pumps, such as syringe pumps,gravity feed, and the like, and are selected by control of valves 1450.Reagents from the fluidics circuit 1402 can be driven to the wastecontainers 1420 and 1436. The control system 1418 includes controllersfor valves 1450 that generate signals for opening and closing via anelectrical connection 1416.

The control system 1418 also includes controllers for other componentsof the system, such as a wash solution valve 1424 connected thereto bythe electrical connection 1422, and the reference electrode 1428. Thecontrol system 1418 can also include control and data acquisitionfunctions for the biosensor 1434. In one mode of operation, the fluidiccircuit 1402 delivers a sequence of selected reagents 1, 2, 3, 4, or 5to the biosensor 1434 under programmed control of the control system1418, such that in between selected reagent flows, the fluidics circuit1402 is primed and washed with a wash solution 1426, and the biosensor1434 is washed with the wash solution 1426. Fluids entering thebiosensor 1434 exit through the outlet 1440 and are deposited in thewaste container 1436. A similar setup can be used for optical sequencingsystems, with photodiodes or CCD cameras for example.

The enclosure and container components can be constructed from a varietyof materials, including metals, glass, ceramics, polymers, or the like.The compressible member can be formed of any inert material such aspolyurethane and can be a sponge that is compressible.

As mentioned above, fluidic circuits can be fabricated by a variety ofmethods and materials. Factors to be considered in selecting materialsinclude degree of chemical inertness required, operating conditions,e.g. temperature, and the like, volume of reagents to be delivered,whether or not a reference voltage is required, manufacturability, andthe like. For meso-scale and larger scale fluid deliveries, conventionalmilling techniques can be used to fabricate parts that can be assembledinto fluidic circuits of the invention. In one aspect, plastics such aspolycarbonate, polymethyl methacrylate, and the like, can be used tofabricate fluidics circuits of the invention.

The reagent storage devices as described herein can operate as such.Each enclosure can start empty with a container within the enclosure.The container can contain liquid nucleotides or lyophilized nucleotides.The enclosure is deflated by removing any air within the enclosure. Thiscan be done by either suctioning any air out of the bag, by pressurizingthe area surrounding the enclosure, or by adding liquid to the enclosuredriving any air out. In some instances, during operation, an additionalsolution can be introduced to the enclosure and the amount introducedcan be limited by the walls of the cavity surrounding the enclosure orwhen pressure in the cavity is equal to or exceeds the pressure of theenclosure during expansion. Any bubbles introduced to the system can bepurged from the enclosure. Mixing of the reagents or solutions in thebag can occur by pressurizing and depressurizing the enclosure.Alternatively the solutions can be mixed by vibrating the body in theenclosure. In some embodiments, mixing can occur with pressurization ofthe contents of the enclosure. For example, external gas pressure can beapplied to the bag, causing internal solutions to flow into thecontainer when the compressible volume or member compresses. As the gaspressure is quickly released, the pressure ‘charged’ within thecompressible volume or member forces solution out through passages inthe container thereby mixing liquid.

In particular, the above described devices and methods provide technicaladvantages that address transportation and storage issues for sensitivereagents, such as biomolecules, each, nucleotides. Transportation ofsuch reagents can be accomplished with less concern of contamination, pHchange, and degradation.

Methods and devices for mixing a reagent, such as methods and devicesfor mixing a reagent and fluid within an enclosure, are described. Invarious implementations, the methods or devices can be combined with abiosensor in fluidic communication with the mixed reagent solution toprovide reliable measurement results. The methods or devices areexemplified in a number of implementations, some of which are summarizedthroughout the specification.

In one aspect, an apparatus includes an enclosure and a containerdisposed within the enclosure. The container defines an internal cavityand defines a passage providing fluidic communication between theinternal cavity and the exterior of the container. A compressible memberis disposed within the internal cavity. A reagent is disposed within theinternal cavity.

In a related aspect, the enclosure is a flexible bag enclosure. Theenclosure includes a fluid port to provide fluid access to the interiorof the enclosure.

In a related aspect, an arm is coupled to the container. A set of armscan be coupled to the container to position the container approximatelycentrally within the enclosure. An arm can be coupled to the containerand the fluid port to position the container approximately centrallywithin the enclosure. The container can include a cap to form theinternal cavity. A flange can be coupled to the container and the fluidport to orient the container within the enclosure. The reagent comprisesa lyophilized nucleotide or an analog thereof.

In a related aspect, the apparatus further includes a case defining avolume, the enclosure disposed within the volume. The case includes agas inlet port providing access to the volume and external to theenclosure. The case includes a lid to secure the enclosure and providefluidic access to a fluid port of the enclosure.

In another aspect, an apparatus includes a case defining a volume andhaving a gas port. A plurality of enclosures are disposed within thevolume and secured to the case. Each enclosure of the plurality ofenclosures includes a fluid port providing fluidic access to theinterior of the enclosure. A container is disposed within eachenclosure. The container defines an internal cavity and defines apassage providing fluidic communication between the internal cavity andthe exterior of the container. A compressible member is disposed withinthe internal cavity. A reagent is disposed within the internal cavity.

In a related aspect, each enclosure of the plurality of enclosures issecured to the container. The container provides fluidic access to thefluid port of the each enclosure. The enclosure can be a flexible bagenclosure. Each enclosure further includes a set of flexible armscoupled to the container to position the container approximatelycentrally within the enclosure. The reagent of the each enclosureincludes a unique lyophilized nucleotide or an analog thereof. Thecontainer can include a cap to form the internal cavity. A flange can becoupled to the container and the fluid port to orient the containerwithin the enclosure.

In another aspect, a method for preparing a reagent solution includesfilling an enclosure with a fluid through a fluid port of the enclosure.The enclosure includes a container disposed within the enclosure. Thecontainer defines an internal cavity and defines a passage providingfluidic communication between the internal cavity and the exterior ofthe container. A compressible member is disposed within the internalcavity. A reagent is disposed within the internal cavity. The fluidwithin the enclosure is pressurized, the fluid flowing into the internalcavity of the container and compressing the compressible member. Thefluid within the enclosure is depressurized, the compressible memberdecompressing and ejecting the fluid and reagent from the internalcavity. The reagent mixes with the fluid.

In a related aspect, the enclosure is a flexible enclosure. Pressurizingincludes increasing a gas pressure external to the flexible enclosure.

In a related aspect, the enclosure is disposed within a case.Pressurizing includes increasing a gas pressure within the case andexternal to the enclosure.

In a related aspect, the pressurizing and the depressurizing steps arerepeated, and the reagent is further mixed with the fluid. The fluidflows into the internal cavity of the container and compresses thecompressible member until the pressure within the internal cavity andexerted on the compressible member is approximately equal to a pressurewithin the enclosure and external to the container. Upon depressurizing,a pressure within the internal cavity as imposed by the compressiblemember is greater than a pressure within the enclosure and external tothe container. The fluid and the reagent eject from the internal cavitythrough the passage until the pressure within the internal cavity isapproximately equal to a pressure within the enclosure and external tothe container. The fluid and the reagent are released through the fluidport.

In another aspect, a method of preparing a reagent solution includesinserting a container into an enclosure. The container defines aninternal cavity and defines a passage providing fluidic communicationbetween the internal cavity and the exterior of the container. Thecontainer includes a compressible member disposed within the internalcavity and a reagent disposed within the internal cavity. The containeris sealed within the enclosure with a fluid port. The enclosure isfilled with a fluid through the fluid port. The fluid within theenclosure is pressurized, the fluid flowing into the internal cavity ofthe container and compressing the compressible member. The fluid withinthe enclosure is depressurized, the compressible member decompressingand ejecting the fluid and reagent from the internal cavity. The reagentmixes with the fluid.

In a related aspect, the enclosure is a flexible enclosure. Pressurizingincludes increasing a gas pressure external to the flexible enclosure.

In a related aspect, the enclosure is disposed within a case.Pressurizing includes increasing a gas pressure within the case andexternal to the enclosure.

In a related aspect, the pressurizing and the depressurizing steps arerepeated, and the reagent is further mixed with the fluid. The fluidflows into the internal cavity of the container and compresses thecompressible member until the pressure within the internal cavity andexerted on the compressible member is approximately equal to a pressurewithin the enclosure and external to the container. Upon depressurizing,a pressure within the internal cavity as imposed by the compressiblemember is greater than a pressure within the enclosure and external tothe container. The fluid and the reagent eject from the internal cavitythrough the passage until the pressure within the internal cavity isapproximately equal to a pressure within the enclosure and external tothe container. The fluid and the reagent are released through the fluidport.

In another aspect, a method of preparing a reagent solution includessecuring a plurality of enclosures within a volume of a case. The caseincludes a gas port. Each enclosure includes a container disposed withinthe enclosure. The container defines an internal cavity and defines apassage providing fluidic communication between the internal cavity andthe exterior of the container. The container includes a compressiblemember disposed within the internal cavity and a reagent disposed withinthe internal cavity. The enclosure is filled with a fluid through afluid port of the enclosure. A gas pressure of the volume of the case isincreased via the gas port to pressurize the fluid within the enclosure.The fluid flows into the internal cavity of the container and compressesthe compressible member. The gas pressure of the volume of the case isdecreased via the gas port to depressurize the fluid within theenclosure. The compressible member decompresses and ejects the fluid andreagent from the internal cavity. The reagent mixes with the fluid.

In a related aspect, the enclosure is a flexible enclosure. Pressurizingincludes increasing a gas pressure external to the flexible enclosure.The pressurizing and the depressurizing steps are repeated, and thereagent is further mixed with the fluid. The fluid flows into theinternal cavity of the container and compresses the compressible memberuntil the pressure within the internal cavity and exerted on thecompressible member is approximately equal to a pressure within theenclosure and external to the container. Upon depressurizing, a pressurewithin the internal cavity as imposed by the compressible member isgreater than a pressure within the enclosure and external to thecontainer. The fluid and the reagent eject from the internal cavitythrough the passage until the pressure within the internal cavity isapproximately equal to a pressure within the enclosure and external tothe container. The fluid and the reagent are released through the fluidport.

In a first aspect, a method of preparing a reagent solution includesproviding a plurality of enclosures within a volume of a cartridge. Thecartridge includes a gas port and the enclosure. The enclosure includesa container disposed within the enclosure. The container defines aninternal cavity and defines a passage providing fluidic communicationbetween the internal cavity and the exterior of the container. Theenclosure also includes a reagent disposed within the internal cavity.The method further includes filling the enclosure with a fluid through afluid port of the enclosure; increasing a gas pressure of the volume ofthe cartridge via the gas port to pressurize the fluid within theenclosure, the fluid flowing into the internal cavity of the containerand compressing the compressible volume; and decreasing the gas pressureof the volume of the cartridge via the gas port to depressurize thefluid within the enclosure, the compressible volume decompressing andejecting the fluid and reagent from the internal cavity, the reagentmixing with the fluid.

In an example of the first aspect, the enclosure further includes acompressible member disposed within the internal cavity.

In another example of the first aspect and the above examples, theenclosure is a flexible enclosure and wherein pressurizing includesincreasing a gas pressure external to the flexible enclosure.

In a further example of the first aspect and the above examples, themethod further includes repeating pressurizing and depressurizing,wherein the reagent is further mixed with the fluid.

In an additional example of the first aspect and the above examples, thefluid flows into the internal cavity of the container and compresses thecompressible volume until the pressure within the internal cavity andexerted on the compressible volume is approximately equal to a pressurewithin the enclosure and external to the container.

In another example of the first aspect and the above examples, upondepressurizing, a pressure within the internal cavity as imposed by thecompressible volume is greater than a pressure within the enclosure andexternal to the container, the fluid and the reagent eject from theinternal cavity through the passage until the pressure within theinternal cavity is approximately equal to a pressure within theenclosure and external to the container.

In a further example of the first aspect and the above examples, themethod of releasing the fluid and the reagent through the fluid port.

In a second aspect, an apparatus includes an enclosure; a containerdisposed within the enclosure, the container defining an internal cavityhaving a compressible volume and defining a passage providing fluidiccommunication between the internal cavity and the exterior of thecontainer; and a reagent disposed within the internal cavity.

In an example of the second aspect, the apparatus further includes acompressible member disposed within the internal cavity.

In another example of the second aspect and the above examples, thecompressible member includes a resilient foam.

In a further example of the second aspect and the above examples, theenclosure is a flexible enclosure.

In an additional example of the second aspect and the above examples,the enclosure includes a fluid port to provide fluid access to theinterior of the enclosure.

In another example of the second aspect and the above examples, theapparatus further includes an arm coupled to the container.

In a further example of the second aspect and the above examples, theapparatus further includes a set of arms coupled to the container toposition the container approximately centrally within the enclosure.

In an additional example of the second aspect and the above examples,the apparatus further includes an arm coupled to the container and thefluid port to position the container approximately centrally within theenclosure.

In another example of the second aspect and the above examples, thecontainer includes an insert coupled to a fitting to form the internalcavity.

In a further example of the second aspect and the above examples, theapparatus further includes a flange coupled to the container.

In an additional example of the second aspect and the above examples,the reagent comprises a lyophilized nucleotide or an analog thereof, anucleotide solution, or a pH-adjusting solution.

In another example of the second aspect and the above examples, theapparatus further includes a cartridge defining a volume, the enclosuredisposed within the volume.

In a further example of the second aspect and the above examples, thecartridge includes a gas inlet port providing access to the volume andexternal to the enclosure.

In an additional example of the second aspect and the above examples,the cartridge includes a lid to secure the enclosure and provide fluidicaccess to a fluid port of the enclosure.

In a third aspect, an apparatus includes a case defining a volume andhaving a gas port; and a plurality of enclosures disposed within thevolume and secured to the case. Each enclosure of the plurality ofenclosures includes a fluid port providing fluidic access to theinterior of the enclosure; a container disposed within the eachenclosure, the container defining an internal cavity having acompressible volume and defining a passage providing fluidiccommunication between the internal cavity and the exterior of thecontainer; and a reagent disposed within the internal cavity.

In an example of the third aspect, the apparatus further includes acompressible member disposed within the internal cavity. For example,the compressible member includes a resilient foam.

In another example of the third aspect and the above examples, the eachenclosure of the plurality of enclosures is secured to the container,the container providing fluidic access to the fluid port of the eachenclosure.

In a further example of the third aspect and the above examples, thecontainer is secured to the enclosure through an arm coupling thecontainer to the enclosure.

In an additional example of the third aspect and the above examples, theenclosure is a bag enclosure.

In another example of the third aspect and the above examples, the eachenclosure further includes a set of arms coupled to the container toposition the container approximately centrally within the enclosure.

In a further example of the third aspect and the above examples, thereagent of the each enclosure includes a unique lyophilized nucleotideor an analog thereof, a nucleotide solution, or a pH-adjusting solution.

In an additional example of the third aspect and the above examples, thecontainer includes an insert coupled to a fitting to form the internalcavity.

In another example of the third aspect and the above examples, theapparatus further includes a flange coupled to the container and thefluid port.

In a fourth aspect, a method for preparing a reagent solution includesfilling an enclosure with a fluid through a fluid port of the enclosure.The enclosure includes a container disposed within the enclosure, thecontainer defining an internal cavity having a compressible volume anddefining a passage providing fluidic communication between the internalcavity and the exterior of the container; and a reagent disposed withinthe internal cavity. The method further includes pressurizing the fluidwithin the enclosure, the fluid flowing into the internal cavity of thecontainer and compressing the compressible member; and depressurizingthe fluid within the enclosure, the compressible member decompressingand ejecting the fluid and reagent from the internal cavity, the reagentmixing with the fluid.

In an example of the fourth aspect, the container further includes acompressible member disposed within the internal cavity.

In another example of the fourth aspect and the above examples, theenclosure is a flexible enclosure and wherein pressurizing includesincreasing a gas pressure external to the flexible enclosure.

In a further example of the fourth aspect and the above examples, theenclosure is disposed within a case, wherein pressurizing includesincreasing a gas pressure within the case and external to the enclosure.

In an additional example of the fourth aspect and the above examples,the method further includes repeating pressurizing and depressurizing,wherein the reagent is further mixed with the fluid.

In a further example of the fourth aspect and the above examples, thefluid flows into the internal cavity of the container and compresses thecompressible member until the pressure within the internal cavity andexerted on the compressible member is approximately equal to a pressurewithin the enclosure and external to the container. In an additionalexample, wherein upon depressurizing, a pressure within the internalcavity as imposed by the compressible member is greater than a pressurewithin the enclosure and external to the container, the fluid and thereagent eject from the internal cavity through the passage until thepressure within the internal cavity is approximately equal to a pressurewithin the enclosure and external to the container.

In an additional example of the fourth aspect and the above examples,the method further includes releasing the fluid and the reagent throughthe fluid port.

In a fifth aspect, a method of preparing a reagent solution includesinserting a container into an enclosure, the container defining aninternal cavity and defining a passage providing fluidic communicationbetween the internal cavity and the exterior of the container. Thecontainer includes a compressible member disposed within the internalcavity and a reagent disposed within the internal cavity. The methodalso includes sealing the container within the enclosure with a fluidport; filling the enclosure with a fluid through the fluid port;pressurizing the fluid within the enclosure, the fluid flowing into theinternal cavity of the container and compressing the compressiblemember; and depressurizing the fluid within the enclosure, thecompressible member decompressing and ejecting the fluid and reagentfrom the internal cavity, the reagent mixing with the fluid.

In an example of the fifth aspect, the enclosure is a flexible enclosureand wherein pressurizing includes increasing a gas pressure external tothe flexible enclosure.

In another example of the fifth aspect and the above examples, theenclosure is disposed within a case, wherein pressurizing includesincreasing a gas pressure within the case and external to the enclosure.

In a further example of the fifth aspect and the above examples, themethod further includes repeating pressurizing and depressurizing,wherein the reagent is further mixed with the fluid.

In an additional example of the fifth aspect and the above examples, thefluid flows into the internal cavity of the container and compresses thecompressible member until the pressure within the internal cavity andexerted on the compressible member is approximately equal to a pressurewithin the enclosure and external to the container.

In another example of the fifth aspect and the above examples, whereinupon depressurizing, a pressure within the internal cavity as imposed bythe compressible member is greater than a pressure within the enclosureand external to the container, the fluid and the reagent eject from theinternal cavity through the passage until the pressure within theinternal cavity is approximately equal to a pressure within theenclosure and external to the container.

In a further example of the fifth aspect and the above examples, themethod further includes releasing the fluid and the reagent through thefluid port.

Note that not all of the activities described above in the generaldescription or the examples are required, that a portion of a specificactivity may not be required, and that one or more further activitiesmay be performed in addition to those described. Still further, theorder in which activities are listed are not necessarily the order inwhich they are performed.

In the foregoing specification, the concepts have been described withreference to specific embodiments. However, one of ordinary skill in theart appreciates that various modifications and changes can be madewithout departing from the scope of the invention as set forth in theclaims below. Accordingly, the specification and figures are to beregarded in an illustrative rather than a restrictive sense, and allsuch modifications are intended to be included within the scope ofinvention.

As used herein, the terms “comprises,” “comprising,” “includes,”“including,” “has,” “having” or any other variation thereof, areintended to cover a non-exclusive inclusion. For example, a process,method, article, or apparatus that comprises a list of features is notnecessarily limited only to those features but may include otherfeatures not expressly listed or inherent to such process, method,article, or apparatus. Further, unless expressly stated to the contrary,“or” refers to an inclusive-or and not to an exclusive-or. For example,a condition A or B is satisfied by any one of the following: A is true(or present) and B is false (or not present), A is false (or notpresent) and B is true (or present), and both A and B are true (orpresent).

Also, the use of “a” or “an” are employed to describe elements andcomponents described herein. This is done merely for convenience and togive a general sense of the scope of the invention. This descriptionshould be read to include one or at least one and the singular alsoincludes the plural unless it is obvious that it is meant otherwise.

Benefits, other advantages, and solutions to problems have beendescribed above with regard to specific embodiments. However, thebenefits, advantages, solutions to problems, and any feature(s) that maycause any benefit, advantage, or solution to occur or become morepronounced are not to be construed as a critical, required, or essentialfeature of any or all the claims.

After reading the specification, skilled artisans will appreciate thatcertain features are, for clarity, described herein in the context ofseparate embodiments, may also be provided in combination in a singleembodiment. Conversely, various features that are, for brevity,described in the context of a single embodiment, may also be providedseparately or in any subcombination. Further, references to valuesstated in ranges include each and every value within that range.

What is claimed is:
 1. An apparatus comprising: a case defining a volumeand having a gas port; and a plurality of enclosures disposed within thevolume and secured to the case, each enclosure of the plurality ofenclosures comprising: a fluid port providing fluidic access to theinterior of the enclosure; a container disposed within the eachenclosure, the container defining an internal cavity having acompressible volume and defining a passage providing fluidiccommunication between the internal cavity and the exterior of thecontainer, wherein the container includes an insert coupled to a fittingto form the internal cavity; and a reagent disposed within the internalcavity.
 2. The apparatus of claim 1, further comprising a compressiblemember disposed within the internal cavity.
 3. The apparatus of claim 2,wherein the compressible member includes a resilient foam.
 4. Theapparatus of claim 1, wherein the each enclosure of the plurality ofenclosures is secured to the container, the container providing fluidicaccess to the fluid port of the each enclosure.
 5. The apparatus ofclaim 1, wherein the container is secured to the enclosure through anarm coupling the container to the enclosure.
 6. The apparatus of claim1, wherein the enclosure is a bag enclosure.
 7. The apparatus of claim1, wherein the each enclosure further includes a set of arms coupled tothe container to position the container approximately centrally withinthe enclosure.
 8. The apparatus of claim 1, wherein the reagent of theeach enclosure includes a unique lyophilized nucleotide or an analogthereof, a nucleotide solution, or a pH-adjusting solution.
 9. Theapparatus claim 1, further comprising a flange coupled to the containerand the fluid port.